LOCUS       sequence-382350-        3840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression vector for  a low-affinity calcium indicator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3840)
  AUTHORS   Valiente-Gabioud AA, Garteizgogeascoa Suner I, Idziak A, Fabritius 
            A, Basquin J, Angibaud J, Nagerl UV, Singh SP, Griesbeck O
  TITLE     Fluorescent sensors for imaging of interstitial calcium.
  JOURNAL   Nat Commun. 2023 Oct 5;14(1):6220. doi: 10.1038/s41467-023-41928-w.
  PUBMED    37798285
REFERENCE   2  (bases 1 to 3840)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2023-10-5"; volume: "14(1):6220. doi"; pages: " 
            10.1038/s41467-023-41928-w"
COMMENT     Sequence Label: GreenT-EC.c - pRSET
FEATURES             Location/Qualifiers
     source          1..3840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     95..114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     348..365
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     450..469
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        450..468
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             500..522
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             542..571
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             578..610
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(614..622,623..637)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      644..653
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1449..1458
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1604..1621)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1671..1689)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1685..1732
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1829..2284
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1916..1935)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2126..2147
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2310..2414
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2415..2483,2484..3275)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2633..2652)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3446..3840,1..194)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
       61 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      121 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      181 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      241 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
      301 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
      361 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
      421 attaatgcag gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg
      481 gtttccctct agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc
      541 tcatcatcat catcatcacc atcaccatca ccgcggtatg gctagcatga ctggtggaca
      601 gcaaatgggt cgggatctgt acgacgatga cgataaggat ccggccacca tggtctccaa
      661 gggggaggag gacaacatgg ggagcctgcc agctacccac gagctgcata tctttggatc
      721 tattaacggg gtggatttcg acatggttgg acaaggtagc gggaatccta atgatgggta
      781 tgaagagctt aaccttaaaa gcactaaggg cgatctgcaa ttttcacctt ggattctcgt
      841 cccccacatt gggtatgggt tccatcaata tctcccttac ccggatggaa tgtctccctt
      901 ccaagcggct atggtagatg gtagtggtta ccaagttcac aggactatgc aattcgaaga
      961 cggcgcctca ttgactgtga actaccgata cacttacgaa ggatctcaca taaaaggtga
     1021 agcgcaggtg aagggcacgg gatttccagc agatgggccg gtaatgacta actcacttac
     1081 tgcggcagac ttggggtggg attcagaaga ggaactttcc gaatgtttta gaatttttga
     1141 ttttgacggt aacggattca ttgaccgcga agagttcggt gacataatca gattgactgg
     1201 ggagcaactt acggatgagg acgtagacga aatttttggc gactcagata cggataagaa
     1261 ttcaggccgg atcgactttg atgaatttct gaaaatggtc gaaaatgttc agctcacgga
     1321 caaccgaaca acgaaaaaga cctatccaaa tgacaagaca attatctcaa ctttgaaatg
     1381 gagctacacc acggacaacg gcaagaggta tcggtccacc gctatcacta tttatacatt
     1441 tgctaaaccc atggcggcta actacctcaa gaatcaacct atgtacgtgt ttcgcaagac
     1501 tgagcttaaa catagcaaaa ccgaattgaa tttcaaggag tggcagaagg cgtttacgga
     1561 cgtaatgggg atggatgagc tttataagtg ataataggaa ttcctatagt gtcacctaaa
     1621 tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     1681 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     1741 aggaactata tccggatctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     1801 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     1861 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     1921 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     1981 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2041 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2101 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2161 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2221 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     2281 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2341 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2401 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2461 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2521 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2581 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2641 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2701 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2761 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2821 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2881 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2941 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3001 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3061 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3121 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3181 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3241 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3301 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3361 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3421 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3481 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3541 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3601 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3661 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3721 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3781 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
//
